从炎症免疫角度探究心阳片改善尿毒症心肌病的作用和机制  

Exploring the role and mechanism of Xinyang Tablet in improving uremic cardiomyopathy from the perspective of inflammation and immunity

作　　者：倪世豪 刘东华 何星灵 欧阳小露 张小娇 李锦 陈星伶 陈梓欣[1] 王陵军[1] 冼绍祥[1] 汪朝晖[1] 廖慧丽[1] 鲁路 杨忠奇[1] NI Shihao;LIU Donghua;HE Xingling;OUYANG Xiaolu;ZHANG Xiaojiao;LI Jin;CHEN Xingling;CHEN Zixin;WANG Lingjun;XIAN Shaoxiang;WANG Zhaohui;LIAO Huili;LU Lu;YANG Zhongqi(State key laboratory of traditional Chinese medicine syndrome,The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China)

机构地区：[1]中医证候全国重点实验室、广州中医药大学第一附属医院,510405

出　　处：《环球中医药》2024年第7期1267-1273,共7页Global Traditional Chinese Medicine

基　　金：国家中医临床研究基地建设项目(国中医药科技函〔2018〕131号);国家自然科学基金(82374406);青年人才托举工程项目(2021-QNRC2-B30);广东省自然科学基金(2021A1515011457);广州市科技计划项目重点研发计划(202206080015);广州中医药大学青年创新拔尖人才团队培育揭榜挂帅项目;2023年度广州中医药大学第一临床医学院优秀博士学位论文培育项目(YB202301);广州市市校(院)联合资助项目基础与应用基础研究项目(202201020471)。

摘　　要：目的从炎症免疫角度探究心阳片对尿毒症心肌病的治疗作用及其机制。方法采用5/6肾切除术建立尿毒症心肌病小鼠模型,随机分为模型组和心阳片组,每组15只;另设15只假手术小鼠作为假手术组。心阳片组术后给予心阳片0.34 g/(kg·d)灌胃8周,假手术组和模型组给予等量0.9%氯化钠溶液灌胃8周。通过心脏转录组数据结合生物信息学手段分析尿毒症心肌病小鼠的心脏炎症免疫状态及心阳片的调节效果和作用机制。通过实时荧光定量PCR(Real-time Quantitative PCR,qPCR)法检测各组小鼠心脏中炎症因子和炎症免疫细胞相关标记物的mRNA表达水平。通过免疫荧光法观察各组小鼠心脏中炎症免疫细胞的浸润情况。结果心脏转录组GO富集分析结果显示,心阳片可能参与抑制尿毒症心肌病的多个炎症免疫相关生物过程。心脏转录组分析结果显示,心阳片抑制了心脏炎症因子(白细胞介素-1β、白细胞介素-6、白细胞介素-18、肿瘤坏死因子)和炎症免疫细胞相关标记物(CD_(86)、CD_(72)、淋巴细胞抗原6c1、整合素β2)水平,且减少了炎症免疫细胞(中性粒细胞、单核细胞、CD_(86)+巨噬细胞、CD_(72)hi巨噬细胞)的浸润。qPCR结果显示,与模型组相比,心阳片组小鼠心脏中白细胞介素-1β、白细胞介素-6、白细胞介素-18、肿瘤坏死因子mRNA水平均显著下调(P<0.05)。免疫荧光结果显示,与模型组相比,心阳片组中心脏中性粒细胞、单核细胞、CD_(72)hi巨噬细胞的浸润均显著减少。此外,基因集富集分析结果显示,心阳片组核因子-κB、c-Jun氨基末端激酶、细胞外调节蛋白激酶1/2和p38丝裂原活化蛋白激酶信号通路被明显抑制(富集评分分别为-0.55、-0.54、-0.43、-0.50)。结论心阳片可能通过抑制心脏的炎症免疫反应以改善尿毒症心肌病。Objective This study aimed to explore the therapeutic effect and mechanism of Xinyang Tablet on uremic cardiomyopathy from the perspective of inflammation-immunity.Methods A mouse model of uremic cardiomyopathy was established by 5/6 nephrectomy,and randomly divided into the model group and Xinyang Tablet group,with 15 mice in each group;another 15 sham-operated mice were used as the sham group.The Xinyang Tablet group was given Xinyang tablet(0.34 g/kg)by gavage after surgery,while the sham group and the model group were given an equal amount of 0.9%sodium chloride solution by gavage,for 8 weeks.The cardiac inflammation-immunity status and the regulatory effect and mechanism of Xinyang Tablet on uremic cardiomyopathy mice were analyzed by combining cardiac transcriptome data with bioinformatics methods.The mRNA expression levels of inflammatory factors and inflammation-immune cell-related markers in the hearts of mice in each group were detected by Real-time Quantitative PCR(qPCR).The infiltration of inflammatory immune cells in the hearts of mice in each group was observed by the immunofluorescence method.Results GO enrichment analysis of cardiac transcriptome showed that Xinyang Tablet might inhibit multiple inflammation-related biological processes in the hearts of uremic cardiomyopathy mice.Cardiac transcriptome analysis showed that Xinyang tablet inhibited the levels of cardiac inflammatory factors(IL-1β、IL-6、IL-18、TNF)and inflammation-immune cell-related markers(CD_(86),CD_(72),Ly6c1,Itgb2),and reduced the infiltration of inflammation-immune cells(neutrophils,monocytes,CD_(86)+macrophages,CD_(72)hi macrophages).The results of qPCR showed that the mRNA levels of IL-1β,IL-6,IL-18 and TNF in the hearts of the Xinyang Tablet group were significantly down-regulated compared with those of model group(P<0.05).Immunofluorescence results showed that the infiltration of neutrophils,monocytes and CD_(72)hi macrophages in the hearts of the Xinyang Tablet group were significantly reduced compared with those of

关 键 词：尿毒症心肌病 心阳片 炎症免疫反应 免疫细胞 

分 类 号：R285.5[医药卫生—中药学]